PEROXIDASE EXTRACTION AND PURIFICATION FROM WATER MIMOSA

Abstract

Abstract

Peroxidase is an enzyme used extensively in a variety of fields including medicine, agriculture and industry. It can be found in animals, plants and microorganisms. At present, the application of peroxidase is from horseradish roots, which are imported from abroad at a high cost. The objective of this research was to extract peroxidase from 4 parts (leaf, stem, white tissue, and root) of the water mimosa (Neptunia oleracea Lour), a local Thai vegetable. Peroxidase was extracted by using 20 mM Tris-HCl buffer, and pH 7.2 at 4 °C. Subsequently, the enzyme was purified by using NaCl concentrations of 0.0 M and 0.1 M in
20 mM Tris-HCl pH 7.2 with a DEAE-Sepharose column. The molecular weight was calculated with the
SDS-PAGE method. The study results showed that peroxidase activity was found only in the leaf and stem of water mimosa with the ideal total enzyme activity of peroxidase at 0.0 M NaCl. The molecular weight of peroxidase in a single subunit was approximately 32.3 ± 2 kDa (n=5), and the perfect total enzyme activity was found. In comparison with the enzymes from rambutan (Nephelium lappaceum Linn), popping pod (Ruellia tuberosa), and water morning glory (Ipomoea aquatica Ipomoea aquatica) at an equal amount of 5 grams, as in previous publications at 50-5,000 folds, the peroxidase enzyme extracted from water mimosa was revealed to possess the “perfect total activity enzyme”.